Basic Colorimetric Proliferation Assays: MTT, WST, and Resazurin

Methods Mol Biol. 2017:1601:1-17. doi: 10.1007/978-1-4939-6960-9_1.

Abstract

This chapter describes selected assays for the evaluation of cellular viability and proliferation of cell cultures. The underlying principle of these assays is the measurement of a biochemical marker to evaluate the cell's metabolic activity. The formation of the omnipresent reducing agents NADH and NADPH is used as a marker for metabolic activity in the following assays. Using NADH and NADPH as electron sources, specific dyes are biochemically reduced which results in a color change that can be determined with basic photometrical methods. The assays selected for this chapter include MTT, WST, and resazurin. They are applicable for adherent or suspended cell lines, easy to perform, and comparably economical. Detailed protocols and notes for easier handling and avoiding pitfalls are enclosed to each assay.

Keywords: Colorimetric proliferation assay; MTT; Metabolic assay; Resazurin; Tetrazolium salts; Viability assay; WST.

MeSH terms

  • Biological Assay
  • Calibration
  • Cell Count / methods*
  • Cell Proliferation*
  • Cell Survival*
  • Colorimetry / methods*
  • HeLa Cells
  • Humans
  • Indicators and Reagents / chemistry*
  • NAD / analysis
  • NADP / analysis
  • Oxazines / chemistry*
  • Tetrazolium Salts / chemistry*
  • Thiazoles / chemistry*
  • Xanthenes / chemistry*

Substances

  • 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium monosodium salt
  • Indicators and Reagents
  • Oxazines
  • Tetrazolium Salts
  • Thiazoles
  • Xanthenes
  • NAD
  • resazurin
  • NADP
  • thiazolyl blue